JP-2022530467-A5 -

Dates

Publication Date

20230509

Application Date

20200425

Description

In another embodiment, the present invention provides a method for producing 1,5-pentanediol, 1,6-hexanediol, adipic acid, or 6-hydroxyhexanoate, comprising culturing a population or recombinant microorganism as disclosed herein under suitable conditions that promote the expression of an exogenous nucleic acid as disclosed herein. In one embodiment, the exogenous nucleic acid is overexpressed compared to a wild-type or unmodified equivalent microorganism. In some embodiments, the method further comprises isolating the 1,5-pentanediol, 1,6-hexanediol, adipic acid, or 6-hydroxyhexanoate from the culture or microorganism. In certain embodiments, for example, the following items are provided: (Item 1) A method comprising contacting a pyruvate and an aliphatic aldehyde with an aldol dehydration product biosynthesis polypeptide to produce an aldol dehydration product, The carbonyl group of the aliphatic aldehyde is not conjugated to an alkenyl group, an alkynyl group, or an aromatic group. The method wherein the aldol dehydration product is a compound comprising an aldehyde group or a ketone group and a double bond conjugated to the aldehyde group or ketone group. (Item 2) The method according to item 1, wherein the aldol dehydration product biosynthesis polypeptide is a hydratase-aldolase or comprises a hydratase-aldolase. (Item 3) The method according to item 1, wherein the aldol dehydration product biosynthesis polypeptide is an enzyme with EC number 4.1.2.45, EC number 4.1.2.34, or EC 4.1.1.4, or an enzyme selected from Tables 1 and 5-8, or the method comprising the enzyme. (Item 4) The method according to item 3, wherein the aldol dehydration product biosynthetic polypeptide is present in a microorganism. (Item 5) A method comprising contacting an alkene with an alkene reduction product biosynthesis polypeptide to produce an alkene reduction product, The aforementioned alkene contains a double bond conjugated to a carbonyl group, The method for reducing the double bond conjugated to the carbonyl group in the alkene to a single bond, thereby yielding an alkene reduction product. (Item 6) The method according to item 5, wherein the alkene is the aldol dehydration product described in item 1. (Item 7) The method according to item 6, wherein the alkene reduction product biosynthetic polypeptide is an enzyme belonging to EC1.6.5, or an enzyme selected from Table 9, or comprises the enzyme. (Item 8) The aliphatic aldehyde has the structure shown in formula A-1 below. R a -L 2 -L 1 -C(O)H A-1 or having a salt thereof, in the formula, R a is R" or -OR", Each of L1 and L2 is independently a covalently bonded or divalently substituted linear or branched C1-20 aliphatic group or C1-20 heteroaliphatic group, wherein one or more of its methylene units are arbitrarily and independently replaced by -C≡C-, -C(R'') 2- , -Cy-, -O-, -S-, -S-S-, -N(R'')-, -C(O)-, -C(S)-, -C(NR'')-, -C(O)N(R'')-, -N(R'')C(O)N(R'')-, -N(R'')C(O)O-, -S(O)-, -S(O) 2- , -S(O) 2N (R'')-, -C(O)S-, or -C(O)O-. -Cy- is a 3- to 20-membered monocyclic, dicyclic, or polycyclic ring with any divalent substituted group, and each monocyclic ring independently has 0 to 5 heteroatoms and is either anyly substituted, saturated, partially saturated, or aromatic, Each R'' is independently -R', -C(O)R', -CO₂R ' , or -SO₂R ' . R' is either hydrogen, or an arbitrarily substituted group selected from a C1-10 aliphatic group, a C1-10 heteroaliphatic group having 1 to 5 heteroatoms , a 6-10 membered aryl ring, a 5-10 membered heteroaryl ring having 1 to 5 heteroatoms, and a 3-10 membered heteroring having 1 to 5 heteroatoms, or The method according to item 4, wherein two or more R' groups, together with an intervening atom, form a optionally substituted 3-20 member monoring, diring, or polyring having 0-5 heteroatoms in addition to the intervening atom, and each monoring is independently a 3-20 member ring that is optionally substituted, saturated, partially saturated, or aromatic, having 0-5 heteroatoms. (Item 9) The aldol dehydration product has the following structure, P-2 R a -L 2 -L 1 -CH=CH-C(O)-C(O)OH P-2 or having a salt thereof, in the formula, R a is R" or -OR", Each of L1 and L2 is independently a covalently bonded or divalently substituted linear or branched C1-20 aliphatic group or C1-20 heteroaliphatic group, wherein one or more of its methylene units are arbitrarily and independently replaced by -C≡C-, -C(R'') 2- , -Cy-, -O-, -S-, -S-S-, -N(R'')-, -C(O)-, -C(S)-, -C(NR'')-, -C(O)N(R'')-, -N(R'')C(O)N(R'')-, -N(R'')C(O)O-, -S(O)-, -S(O) 2- , -S(O) 2N (R'')-, -C(O)S-, or -C(O)O-. -Cy- is a 3- to 20-membered monocyclic, dicyclic, or polycyclic ring with any divalent substituted group, and each monocyclic ring independently has 0 to 5 heteroatoms and is either anyly substituted, saturated, partially saturated, or aromatic, Each R'' is independently -R', -C(O)R', -CO₂R ' , or -SO₂R ' . R' is either hydrogen, or an arbitrarily substituted group selected from a